Virus filtration

Abstract

Provided herein are methods of performing viral filtration on a fluid including a recombinant antibody, and the use of these methods in methods of manufacturing or producing the recombinant antibody.

What is claimed is: 
     
       1. A method of performing viral filtration, the method comprising the steps of:
 (a) adjusting the pH of a fluid comprising a recombinant antibody to between about 5.0 and about 6.7, and adding a stabilizing agent to the fluid in an amount sufficient to yield a final concentration of between about 0.1 mM and about 25 mM stabilizing agent in the fluid; 
 (b) flowing the fluid through a pre-filter, wherein the pre-filter comprises a polyamide membrane; and 
 (c) immediately following step (b), flowing the fluid through a virus filter to produce a filtrate comprising the recombinant antibody, 
 wherein: 
 (i) the stabilizing agent is selected from the group consisting of arginine, alanine, aspartic acid, glutamic acid, leucine, lysine, histidine, glycine, sucrose, trehalose, mannitol, and sorbitol; and 
 (ii) (ii) the recombinant antibody comprises:
 a heavy chain variable region comprising:
 a CDR1 comprising a sequence of SEQ ID NO: 1; 
 a CDR2 comprising a sequence of SEQ ID NO: 2; and 
 a CDR3 comprising a sequence of SEQ ID NO: 3; and 
 
 a light chain variable region comprising
 a CDR1 comprising a sequence of SEQ ID NO: 6; 
 a CDR2 comprising a sequence of SEQ ID NO: 7; and 
 a CDR3 comprising a sequence of SEQ ID NO: 8. 
 
 
 
     
     
       2. The method of  claim 1 , wherein the fluid further comprises between about 5 mM and about 300 mM sodium chloride. 
     
     
       3. The method of  claim 1 , wherein the virus filter comprises a polyethersulfone membrane. 
     
     
       4. The method of  claim 1 , wherein the virus filter comprises a polyvinylidene fluoride (PVDF) membrane. 
     
     
       5. The method of  claim 1 , wherein the virus filter comprises a cuprammonium-regenerated cellulose membrane. 
     
     
       6. The method of  claim 1 , wherein prior to (a), the pH of the fluid is between about 7.4 and about 7.8. 
     
     
       7. The method of  claim 1 , wherein the heavy chain variable domain comprises a sequence of SEQ ID NO: 4. 
     
     
       8. The method of  claim 1 , wherein the recombinant antibody comprises a heavy chain comprising a sequence of SEQ ID NO: 5. 
     
     
       9. The method of  claim 1 , wherein the recombinant antibody comprises a light chain variable region comprising a sequence of SEQ ID NO: 9. 
     
     
       10. The method of  claim 1 , wherein the recombinant antibody comprises a light chain comprising a sequence of SEQ ID NO: 10. 
     
     
       11. The method of  claim 1 , wherein the recombinant antibody comprises a heavy chain variable domain comprises a sequence of SEQ ID NO: 4 and a light chain variable region comprising a sequence of SEQ ID NO: 9. 
     
     
       12. The method of  claim 1 , wherein the recombinant antibody comprises a heavy chain comprising a sequence of SEQ ID NO: 5 and a light chain comprising a sequence of SEQ ID NO: 10. 
     
     
       13. A method of performing viral filtration, the method comprising the steps of:
 (a) adjusting the pH of a fluid comprising a recombinant antibody to between about 5.0 and about 6.7, and adding a stabilizing agent to the fluid in an amount sufficient to yield a final concentration of between about 0.1 mM and about 25 mM stabilizing agent in the fluid; 
 (b) performing ultrafiltration/diafiltration on the fluid; 
 (c) following step (b), flowing the fluid through a pre-filter, wherein the pre-filter comprises a polyamide membrane; and 
 (d) immediately following step (c), flowing the fluid through a virus filter to produce a filtrate comprising the recombinant antibody, 
 wherein:
 (i) the stabilizing agent is selected from the group consisting of arginine, alanine, aspartic 
 acid, glutamic acid, leucine, lysine, histidine, glycine, sucrose, trehalose, mannitol, and sorbitol; 
 (ii) the recombinant antibody comprises: a heavy chain variable region comprising: the CDR1 comprising a sequence of SEQ ID NO: 1, the CDR2 comprising a sequence of SEQ ID NO: 2, and the CDR3 comprising a sequence of SEQ ID NO: 3, and a light chain variable region comprising: a CDR1 comprising a sequence of SEQ ID NO: 6, a CDR2 comprising a sequence of SEQ ID NO: 7, and a CDR3 comprising a sequence of SEQ ID NO: 8.